| Size | Price | Stock | Qty |
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| 1mg |
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| 5mg |
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| 10mg | |||
| Other Sizes |
| Targets |
NEK2 (NIMA-related kinase 2). JH295 is a cysteine-targeted inhibitor that alkylates Cys22 of Nek2. The compound shows no activity against Cdk1, Aurora B, or Plk1 and is a selective Nek2 inhibitor.
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| ln Vitro |
JH295 (compound 16; 0.08–20 μM; 45 minutes; RPMI7951 cells) shows no effect on the C22V mutant [1], but it inhibits WT Nek2 with an IC50 of about 1.3 μM.
JH295 inhibits Nek2 with an IC₅₀ of 770 nM. It inhibits cellular Nek2 via alkylation of Cys22. JH295 shows no activity against mitotic kinases Cdk1, Aurora B, or Plk1. The compound is a click chemistry reagent. |
| ln Vivo |
No detailed in vivo activity data has been published in the available literature. JH295 is primarily characterized as an in vitro tool for studying Nek2 function in mitosis. Further in vivo studies in xenograft tumor models would be needed to evaluate its anti-tumor efficacy.
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| Enzyme Assay |
Nek2 kinase activity assay: Recombinant Nek2 is incubated with varying concentrations of JH295, ATP, and a peptide substrate in kinase assay buffer at 30°C for 60 min. The reaction is terminated, and phosphorylation is detected by a luminescent or fluorescence-based method. IC₅₀ values are calculated from dose-response curves.
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| Cell Assay |
Western Blot Analysis[1]
Cell Types: RPMI7951 Cell Tested Concentrations: 0.08 μM, 0.25 μM, 0.74 μM, 2.2 μM, 6.6 μM, 20 μM Incubation Duration: 45 minutes Experimental Results: Inhibition of WT Nek2 in cells, IC50 is approximately 1.3 μM , while having almost no effect on the C22V mutant. Cellular Nek2 inhibition assay: Cells are treated with JH295 at varying concentrations for specified time periods. Nek2 activity is assessed by immunoprecipitation followed by in vitro kinase assay using a substrate. Alternatively, Nek2 autophosphorylation is measured by Western blot using anti-phospho-Nek2 antibody. |
| Animal Protocol |
No in vivo animal experimental protocols have been published for JH295 in the available literature. In vivo studies would typically involve xenograft tumor models in immunocompromised mice, where JH295 would be administered via intraperitoneal or intravenous routes, followed by tumor volume measurement and assessment of Nek2 inhibition.
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| ADME/Pharmacokinetics |
JH295 has a molecular weight of 320.35 g/mol and molecular formula C₁₈H₁₆N₄O₂. Purity is ≥98%. CAS number is 1311143-71-1. The compound is a click chemistry reagent. Comprehensive PK parameters remain to be characterized.
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| Toxicity/Toxicokinetics |
No toxicity data has been published. JH295 is for research use only. Standard laboratory safety practices should be followed when handling this compound. As a Nek2 inhibitor, potential effects on mitotic progression should be considered.
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| References | |
| Additional Infomation |
JH295 is a potent, irreversible, and selective Nek2 inhibitor with an IC₅₀ of 770 nM. It inhibits Nek2 via alkylation of Cys22 and shows no activity against Cdk1, Aurora B, or Plk1. JH295 is a click chemistry reagent used to study Nek2 function in mitosis. No clinical trials or approved上市 status have been reported.
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| Molecular Formula |
C18H16N4O2
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|---|---|
| Molecular Weight |
320.35
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| Exact Mass |
320.127
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| CAS # |
1311143-71-1
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| Related CAS # |
JH295 hydrate;2714087-26-8
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| PubChem CID |
53262909
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| Appearance |
Yellow to orange solid powder
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| Density |
1.4±0.1 g/cm3
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| Index of Refraction |
1.716
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| LogP |
1.21
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| Hydrogen Bond Donor Count |
3
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| Hydrogen Bond Acceptor Count |
3
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| Rotatable Bond Count |
3
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| Heavy Atom Count |
24
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| Complexity |
605
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| Defined Atom Stereocenter Count |
0
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| SMILES |
CCC1=NC(=C(N1)C)/C=C\2/C3=C(C=CC(=C3)NC(=O)C#C)NC2=O
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| InChi Key |
ORKOHXAJFGRZCL-LCYFTJDESA-N
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| InChi Code |
InChI=1S/C18H16N4O2/c1-4-16-19-10(3)15(21-16)9-13-12-8-11(20-17(23)5-2)6-7-14(12)22-18(13)24/h2,6-9H,4H2,1,3H3,(H,19,21)(H,20,23)(H,22,24)/b13-9-
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| Chemical Name |
N-[(3Z)-3-[(2-ethyl-5-methyl-1H-imidazol-4-yl)methylidene]-2-oxo-1H-indol-5-yl]prop-2-ynamide
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| Synonyms |
JH-295; JH 295; JH295
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| HS Tariff Code |
2934.99.9001
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| Storage |
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month Note: (1). Please store this product in a sealed and protected environment (e.g. under nitrogen), avoid exposure to moisture. (2). This product is not stable in solution, please use freshly prepared working solution for optimal results. |
| Shipping Condition |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
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| Solubility (In Vitro) |
DMSO : ~55 mg/mL (~171.69 mM)
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|---|---|
| Solubility (In Vivo) |
Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples.
Injection Formulations
Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)(e.g. IP/IV/IM/SC) *Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution. Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline) Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil) Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals). View More
Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)] Oral Formulations
Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium) Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals). View More
Oral Formulation 3: Dissolved in PEG400 (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 3.1216 mL | 15.6079 mL | 31.2159 mL | |
| 5 mM | 0.6243 mL | 3.1216 mL | 6.2432 mL | |
| 10 mM | 0.3122 mL | 1.5608 mL | 3.1216 mL |
*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.
Calculation results
Working concentration： mg/mL；
Method for preparing DMSO stock solution： mg drug pre-dissolved in μL DMSO (stock solution concentration mg/mL). Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug.
Method for preparing in vivo formulation:：Take μL DMSO stock solution, next add μL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O，mix and clarify.
(1) Please be sure that the solution is clear before the addition of next solvent. Dissolution methods like vortex, ultrasound or warming and heat may be used to aid dissolving.
(2) Be sure to add the solvent(s) in order.
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